Eluting reagents, methods and kits for isolating DNA

• US 7,790,865 B1
• Filed: 02/02/1999
• Issued: 09/07/2010
• Est. Priority Date: 02/02/1999
• Status: Expired due to Fees

• Current Assignee: QIAGEN North American Holdings Inc. (Qiagen NV)
• Original Assignee: QIAGEN North American Holdings Inc. (Qiagen NV)
• Inventors: Heath, Ellen M., Shuman, Ruth M.
• Primary Examiner(s): Goldberg; Jeanine A
• Application Number:
  US09/241,637
• Time in Patent Office: 4,235 Days
• Field of Search: 435/91.1, 435/184, 435/219, 435/87, 435/259, 536/23.1, 536/127
• US Class Current: 536/23.1
• CPC Class Codes: C12N 15/1006 by means of a solid support carrier, e.g. particles, polymers
• CPC Class Codes: C12Q 1/6806 Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 2527/125 Specific component of sample, medium or buffer
• CPC Class Codes: C12Q 2527/137 Concentration of a component of medium

Abstract

Eluting reagents and methods for isolating DNA from biological materials are provided.

Claims

1. A kit for isolating DNA comprising:

(a) optionally instruction means for isolating substantially pure DNA from a biological sample;

(b) a DNA purifying composition;

(c) a DNA eluting reagent; and

(d) a solid support selected from a group consisting of glass fiber, nylon, polyester, polyethersulfone, polyolefin, polyvinylidene fluoride, and combinations thereof,

wherein the DNA eluting reagent comprises;

(i) a buffer;

(ii) a base;

(iii) a chelating agent; and

(iv) water,

wherein the chelating agent is present in an amount no greater than 0.1 mM based on the total volume of the DNA eluting reagent, the base is present in an amount between 5-8 mM, and the combined amount of buffer, base, and chelating agent is present in an amount no greater than 20 mM based on the total volume of the DNA eluting reagent.

2. The kit of claim 1, further comprising a vessel for holding the solid support.

3. The kit of claim 1, wherein the solid support is treated with a lysing reagent.

4. The kit of claim 1, wherein the solid support is treated with an RNA digesting enzyme.

5. The kit of claim 1, further comprising an RBC Lysis Reagent comprising ammonium chloride, sodium bicarbonate, and EDTA, for use in separating white cells from red blood cells in mammalian whole blood prior to DNA isolation.

6. The kit of claim 1, further comprising:

(a) a cell suspension reagent comprising Tris, EDTA, and sorbitol; and

(b) a lytic enzyme reagent comprising a lytic enzyme, glycerol, Tris, and calcium chloride, wherein the cell suspension reagent and the lytic enzyme reagent are suitable for use in combination to digest cell walls of yeast and Gram-positive bacteria.

7. The kit of claim 1, further comprising a reagent selected from the group consisting of:

(a) an RBC Lysis reagent;

(b) a cell suspension reagent;

(c) a lytic enzyme reagent;

(d) a protein digesting reagent; and

(e) an isotonic solution.

8. The kit of claim 1, wherein the chelating agent is present in the amount of about 0.0001 to about 0.05 mM.

9. The kit of claim 1, wherein the chelating agent is present in the amount of about 0.0001 to about 0.015 mM.

11. A method for isolating DNA using the kit of claim 1, comprising the steps of:

(a) contacting a biological material that contains DNA with the solid support from the kit of claim 1;

(b) treating the biological material that contains DNA with a DNA purifying reagent;

(c) purifying the DNA from the remainder of the biological materials; and

(d) eluting the DNA from the solid support with the DNA eluting reagent from the kit of claim 1.

12. The method of claim 11, wherein the DNA eluting reagent has a pH of at least 7.

13. The method of claim 11, wherein the base is chosen from the group consisting of alkaline metal hydroxides.

14. The method of claim 11, wherein the buffer is present in an amount from 0.001-20 mM based on the total volume of the DNA eluting reagent.

15. The method of claim 11, wherein the buffer is present in an amount from 0.01-15 mM based on the total volume of the DNA eluting reagent.

16. The method of claim 11, wherein the solid support is contained in a vessel, wherein the vessel is selected from a group consisting of centrifuge tubes, spin tubes, syringes, cartridges, chambers, multiple-well plates, test tubes, and combinations thereof.

17. The method of claim 11, wherein the biological material is selected from the group consisting of eukaryotic cells, prokaryotic cells, microbial cells, bacterial cells, plant cells, mycoplasma, protoza, bacteria, fungi, viruses, and lysates and homogenates thereof.

18. The method of claim 11, wherein the biological material is selected from the group consisting of body fluids, body waste products, excretions, and tissues.

19. The method of claim 11, wherein the biological material is selected from the group consisting of environmental samples taken from air, water, sediment and soil.

20. The method of claim 11, wherein the buffer is Tris, the chelating agent is EDTA, and the base is chosen from the group consisting of alkaline metal hydroxides.

21. The method of claim 11, further comprising the step of heating at greater than 60°

C.

10. A kit for isolating DNA comprising:

(a) optionally instruction means for isolating substantially pure DNA from a biological sample;

(b) a DNA purifying composition;

(c) a DNA eluting reagent, and

(d) a solid support selected from a group consisting of glass fiber, nylon, polyester, polyethersulfone, polyolefin, polyvinylidene fluoride, and combinations thereof,

wherein the DNA eluting reagent comprises;

(i) a buffer;

(ii) a base;

(iii) a chelating agent; and

(iv) water,

wherein the chelating agent is present in an amount no greater than 0.1 mM based on the total volume of the DNA eluting reagent, the base is present in an amount between 5-8 mM, and the combined amount of buffer, base, and chelating agent is present in an amount no greater than 20 mM based on the total volume of the DNA eluting reagent, and wherein the DNA eluting reagent contains no greater than 50 mM NaCl.